Structurally modified acellular tissue engineering scaffolds and methods of production

Abstract

Methods are provided for producing a bioscaffold from natural tissues by oxidizing a decellularized tissue to produce a bioscaffold having pores therein. The pore size and porosity is increased to better accommodate intact cells so that live cells can better infiltrate and inhabit the bioscaffold. The bioscaffold may be freeze-dried or lyophilized, sterilized and (optionally) aseptically packaged for subsequent use. A further aspect of the present invention is a bioscaffold produced by the processes described herein. Methods of treatment using the bioscaffold as a graft or as a biomedical implant for implantation are also provided. Also provided are methods of seeding a bioscaffold with mammalian cells, wherein the seeding carried out either in vitro or in vivo, and wherein a bioscaffold produced as described herein is utilized for said seeding.

1. A method of treating a subject in need of a bioscaffold implant, comprising implanting a bioscaffold in said subject in a treatment effective configuration, wherein said bioscaffold is produced by the process of decellularizing a tissue with an oxidant and detergent simultaneously to remove extraneous material and increase the pore size and porosity therein, wherein at least 70% of cells are removed from said tissue upon said decellularizing. 
     
     
       2. The method of  claim 1 , wherein said oxidant is hydrogen peroxide, peracetic acid, or a mixture thereof. 
     
     
       3. The method of  claim 1 , wherein said oxidant is in an aqueous solution at a concentration of between 1 and 32% (w/v). 
     
     
       4. The method of  claim 1 , wherein said oxidant is in an aqueous solution at a concentration of between 3 and 32% (w/v). 
     
     
       5. The method of  claim 1 , wherein said process further comprises the step of washing said bioscaffold after said decellularizing step to remove residual oxidant and detergent. 
     
     
       6. The method of  claim 1 , wherein said process further comprises the step of freeze-drying said scaffold following said washing step. 
     
     
       7. The method of  claim 5 , wherein said process further comprises the step of sterilizing said scaffold following said washing step. 
     
     
       8. The method of  claim 1 , wherein said process further comprises the step of removing adventitia from said tissue prior to said decellularizing step. 
     
     
       9. The method of  claim 8 , wherein said tissue is a blood vessel. 
     
     
       10. The method of  claim 1 , wherein said tissue is selected from the group consisting of skin, muscle, tendon, bone, meniscus, cartilage, intervertebral discs, and ligament tissue. 
     
     
       11. The method of  claim 1 , wherein said tissue is selected from the group consisting of liver, kidney, and pancreas tissue. 
     
     
       12. The method of  claim 1 , wherein said tissue is nerve tissue. 
     
     
       13. The method of  claim 1 , wherein said tissue is cartilage tissue. 
     
     
       14. The method of  claim 1 , wherein at least 80% of cells are removed from said tissue upon said decellularizing. 
     
     
       15. The method of  claim 1 , wherein at least 90% of the cells are removed from said tissue upon said decellularizing. 
     
     
       16. The method of  claim 1 , wherein said tissue is tendon or ligament tissue. 
     
     
       17. The method of  claim 1 , wherein said tissue is meniscus tissue. 
     
     
       18. The method of  claim 1 , wherein said bioscaffold is seeded with cells prior to said implanting. 
     
     
       19. The method of  claim 18 , wherein said cells are autogeneic. 
     
     
       20. A method of treating a subject in need of a tendon or ligament implant, comprising implanting a bioscaffold in said subject in a treatment effective configuration, wherein said bioscaffold is produced by the process of decellularizing a tendon or ligament tissue with an oxidant and detergent simultaneously to remove extraneous material and increase the pore size and porosity therein, wherein at least 70% of cells are removed from said tissue upon said decellularizing. 
     
     
       21. The method of  claim 20 , wherein at least 80% of cells are removed from said tissue upon said decellularizing. 
     
     
       22. The method of  claim 20 , wherein at least 90% of the cells are removed from said tissue upon said decellularizing. 
     
     
       23. The method of  claim 20 , wherein said bioscaffold is seeded with cells prior to said implanting. 
     
     
       24. The method of  claim 23 , wherein said cells are autogeneic. 
     
     
       25. A method of treating a subject in need of a meniscus implant, comprising implanting a bioscaffold in said subject in a treatment effective configuration, wherein said bioscaffold is produced by the process of decellularizing a meniscus tissue with an oxidant and detergent simultaneously to remove extraneous material and increase the pore size and porosity therein, wherein at least 70% of cells are removed from said tissue upon said decellularizing. 
     
     
       26. The method of  claim 25 , wherein at least 80% of cells are removed from said tissue upon said decellularizing. 
     
     
       27. The method of  claim 25 , wherein at least 90% of the cells are removed from said tissue upon said decellularizing. 
     
     
       28. The method of  claim 25 , wherein said bioscaffold is seeded with cells prior to said implanting. 
     
     
       29. The method of  claim 28 , wherein said cells are autogeneic.